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Matrix Biol. 2016 Jan;49:93-105. doi: 10.1016/j.matbio.2015.06.005. Epub 2015 Jul 26.

Biophysical analysis of a lethal laminin alpha-1 mutation reveals altered self-interaction.

Author information

1
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada; School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom. Electronic address: t.r.patel@bham.ac.uk.
2
Institute for Dental Research and Oral Musculoskeletal Biology and Center for Biochemistry, Medical Faculty, University of Cologne, Cologne 50931, Germany.
3
National Center for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom; Functional Nanomaterials Lab, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Saudi Arabia.
4
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada.
5
National Center for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington LE12 5RD, United Kingdom.
6
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland 4072, Australia.
7
School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.
8
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba R3T 2N2, Canada. Electronic address: jorg.stetefeld@ad.umanitoba.ca.

Abstract

Laminins are key basement membrane molecules that influence several biological activities and are linked to a number of diseases. They are secreted as heterotrimeric proteins consisting of one α, one β, and one γ chain, followed by their assembly into a polymer-like sheet at the basement membrane. Using sedimentation velocity, dynamic light scattering, and surface plasmon resonance experiments, we studied self-association of three laminin (LM) N-terminal fragments α-1 (hLM α-1N), α-5 (hLM α-5N) and β-3 (hLM β-3N) originating from the short arms of the human laminin αβγ heterotrimer. Corresponding studies of the hLM α-1N C49S mutant, equivalent to the larval lethal C56S mutant in zebrafish, have shown that this mutation causes enhanced self-association behavior, an observation that provides a plausible explanation for the inability of laminin bearing this mutation to fulfill functional roles in vivo, and hence for the deleterious pathological consequences of the mutation on lens function.

KEYWORDS:

Analytical ultracentrifugation; CD spectroscopy; Dynamic light scattering; Extracellular matrix; Laminin short arms; Protein self-association; Surface plasmon resonance

PMID:
26215696
DOI:
10.1016/j.matbio.2015.06.005
[Indexed for MEDLINE]

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